1. Field of the Invention
The present invention relates to a bioreactor apparatus for producing cells, selectively harvesting cells and/or tissues, and/or studying cells and/or tissues in vitro under different conditions over short or long time periods, and methods for making and using same.
More specifically, the invention relates to a bioreactor apparatus for producing cells, selectively harvesting cells and/or tissues, and/or studying cells and/or tissues in vitro under different conditions over short or long time periods, where the apparatus includes a laminar flow chamber having a plurality of cell proliferation zones, each zone having an associated inlet and outlet. The apparatus also includes a fluid handling system and a gas handling system, where the two systems are adapted to seed or inoculate, feed, gasify, harvest, stress, and/or study cells over short or long time periods. The apparatus can also include a housing containing the chamber and systems, where the housing includes pressure, temperature, humidity, etc., control systems. The apparatus can include components for automated and unmanned cell seeding, cell proliferating, cell stressing, cell harvesting and cell monitoring operations under antiseptic conditions and for post harvesting analysis, the apparatus can be associated with a cell fluid dynamic/analysis system, especially a micro-fluidic cell handling and analyzing system. The invention also relates to methods for making and using same.
2. Description of the Related Art
Reliable and efficient cell production is now the central focus in the growing demand for drug development and testing, the creation of artificial organs, the creation of engineered tissue, the production of active proteins made by genetically engineered cells, the production of hormones, antibodies, or enzymes, the preparation of viral vaccines, and the advanced understanding of biological processes. Additionally, efforts have been directed to the understanding of cell growth and behavior upon exposure to stressors, cell-to-cell biocompatibility, tissue healing processes, cell and tissue toxicity, cell apoptosis, cell and tissue chemotaxis, and cell and tissue mechanotaxis. Some of these studies require the simultaneous production of cell cultures within the same bioreactor and in a parallel fashion.
One of the crucial problems associated with cell production is that it is time consuming, expensive, and requires skilled operators. To overcome these barriers, some attempts have been made to manufacture bioreactors and develop methods for optimizing cell production and for studying cell chemotaxis and mechanotaxis.
U.S. Pat. No. 5,153,131 (Wolf et al.) discloses a suspension cell culture system where a culture chamber is rotatable about a horizontal axis and has a vertical large area oxygen transmissible membrane spaced a distance about 0.25 inches less than 1.0 inches from a facing vertical wall surface for effective transmission of oxygen to cells in suspension in the culture chamber. The facing vertical wall surface can be a dialysis membrane for exchange of fresh nutrient from a dialysis chamber with cell waste product in the culture chamber.
U.S. Pat. No. 5,496,722 (Goodwin et al.) disclosed a normal mammalian tissue and the culturing process has been developed for the three groups of organ, structural and blood tissue. The cells are grown in vitro under microgravity culture conditions and form three dimensional cells aggregates with normal cell function. The microgravity culture conditions may be microgravity or simulated microgravity created in a horizontal rotating wall culture vessel.
U.S. Pat. No. 5,763,279 (Schwarz et al.) disclosed an apparatus and method of use for a new simply constructed bioreactor made at least partially of gas permeable materials. The bioreactor is useful for culturing cells and tissues in suspension in a liquid nutrient medium with minimum turbulence. The bioreactor may include ports for easy access to the vessel culture, allowing the growth substrate to be varied for optimum performance. A method for culturing cells using the disclosed bioreactor is also described.
U.S. Pat. No. 6,867,040 B2 (Helmstetter et al.) disclosed an apparatus and methods directed to a perfusion culture system in which a rotating bioreactor is used to grow cells in a liquid culture medium, while these cells are attached to an adhesive-treated porous surface. As a result of this arrangement and its rotation, the attached cells divide, with one cell remaining attached to the substrate, while the other cell, a newborn cell is released. These newborn cells are of approximately the same age, that are collected upon leaving the bioreactor. The populations of newborn cells collected are of synchronous and are minimally, if at all, disturbed metabolically.
Thus, there is a need in the art for an improved bioreactor apparatus for perfusing cells with a medium; controlling an amount of fresh and/or recirculated medium through inlet valves supplying medium to each laminar zone in the bioreactor apparatus; controlling waste collection through an outlet valve; controlling temperature, gas transport, gas exchange, and humidity; inoculating cells through an inoculation port; sampling/assaying cells, supernatant, and cell products; selectively harvesting cells; and selectively harvesting cell products.